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Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD+ oxidoreductase (decarboxylating)] was purified from herring testicular tissue to a specific activity of 26.4 μmol NADH/min/mg
protein. Herring testicular tissue is one of the most abundant sources of this enzyme. The purification procedure involved
differential centrifugation of mitochondria and then chromatography on DEAE-Sephacel, Red Agarose and Sephacryl S-300. This
enzyme catalyzes the oxidative decarboxylation of malate in the presence of Mn2+ and either NAD or NADP. Under Vmax conditions the ratios for the rate of NAD/NADP reduction was 1.8. A study of the reductive carboxulation reaction indicated
that this enzyme reaction is reversible; at pH 7.0 the reverse reaction exhibited 22% of the activity of forward reaction.
Some kinetic characteristics of the enzyme were determined. ATP was found to be a competitive inhibitor with respect to malate.
Fumarate reversed ATP inhibition. Regulation of NAD(P)-dependent malic enzyme from herring testicular tissue mitochondria
could respond to changing levels of mitochondrial ATP and fumarate in vivo. 相似文献
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Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD+ oxidoreductase (decarboxylating)] was purified from herring skeletal muscle to a specific activity of 8.2 mol/min/mg. The purification procedure involved chromatography on DEAE-cellulose, Red Agarose and a Sephacryl S-300 with a final recovery of 38% of enzyme activity. This enzyme catalyzes the oxidative decarboxylation of malate in the presence of either NAD or NADP in the presence of Mn2+. Some kinetic characteristics of this enzyme were determined. The pH optimum of activity is 7.0. ATP was shown to be a competitive inhibitor with malate. The inhibition by ATP displayed hyperbolic competitive kinetics with a Ki (ATP) of 0.28 mM in the presence of NAD and 0.75 mM in the presence of NADP. Fumarate reversed ATP inhibition.In vivo, regulation of NAD(P)-dependent malic enzyme might respond to changing levels of mitochondrial ATP and fumarate with the enzyme undergoing kinetic activation by an increase in the concentration of mitochondrial fumarate which could reverse enzyme inhibition by ATP. 相似文献
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Steven W. Effler Carol M. Brooks Jeffrey M. Addess Susan M. Doerr Michelle L. Storey Bruce A. Wagner 《Water, air, and soil pollution》1991,55(3-4):427-444
Concentrations of Cl, total ammonia (TNH3), NO3 plus NO2, total P (TP), and soluble reactive P (SRP) were measured at two sites, located 5 km apart, on Ninemile Creek, New York, for a period of more than 8 mo. The sites bound the most recently formed Solvay waste beds, associated with the production of soda ash, that adjoin the creek. Concentrations of Cl and T-NH3 increased on average by factors of 16.1 and 7.6, respectively, over the monitored stream reach. The estimated average loadings of these materials to the stream over this reach were 2.3 × 105 and 1.2 × 102 kg d?1, respectively. These inputs are attributable to the Solvay waste beds. The loading of Cl from this source has not changed significantly over a 4 yr period since the closure of the soda ash manufacturing facility. This is the single largest source of Cl, and the second largest source of T-NH3, to polluted Onondaga Lake. Profiles of Cl in the lake indicated that at times the creek inflow plunges to subsurface layers as a result of its elevated density. This is at least in part a result of the creek's ionic enrichment. The concentration of SRP decreased by a factor of 2.0 on average over the study reach, probably due to adsorption to the CaCO3 deposits that cover the stream bed in this area. However, the TP load from the creek to the lake is not significantly affected by this phenomenon. 相似文献